LOCUS       sequence-81710-e        7162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible lentiviral vector of human ERAS cDNA with 
            excisable insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7162)
  AUTHORS   Rais Y, Zviran A, Geula S, Gafni O, Chomsky E, Viukov S, Mansour AA,
            Caspi I, Krupalnik V, Zerbib M, Maza I, Mor N, Baran D, Weinberger 
            L, Jaitin DA, Lara-Astiaso D, Blecher-Gonen R, Shipony Z, Mukamel Z,
            Hagai T, Gilad S, Amann-Zalcenstein D, Tanay A, Amit I, Novershtern 
            N, Hanna JH
  TITLE     Deterministic direct reprogramming of somatic cells to pluripotency.
  JOURNAL   Nature. 2013 Oct 3;502(7469):65-70. doi: 10.1038/nature12587. Epub 
            2013 Sep 18.
  PUBMED    24048479
REFERENCE   2  (bases 1 to 7162)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature12587"; journalName: "Nature"; date: "2013-10-3- 3"; 
            volume: "502"; issue: "7469"; pages: "65-70"
COMMENT     Sequence Label: FUW-TetO-lox-ERAS
FEATURES             Location/Qualifiers
     source          1..7162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             604..784
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    831..956
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1449..1682
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1867..1911
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2209..2326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2385..2403,2404..2426,2427..2445,2446..2468,2469..2487,
                     2488..2510,2511..2529,2530..2552,2553..2571,2572..2594,
                     2595..2613,2614..2636,2637..2655)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2688..2725
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2722..2746
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    3551..4139
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3604..3624)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4022..4033)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4142..4158)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4143..4159)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    4663..4696
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4710..4890
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4916..4933)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4922..5033
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5179..5767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5259..5278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5938..6729,6730..6798))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6561..6580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6799..6903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6978..6997)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      601 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      661 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      721 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      781 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
      841 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
      901 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
      961 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1021 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1081 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1141 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1201 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1261 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     1321 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1381 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1441 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1501 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1561 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1621 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1681 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1741 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     1801 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     1861 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     1921 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     1981 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2041 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2101 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2161 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2221 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2281 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2341 ggacagcaga gatccagttt ggttaatgat ctcgagttta ccactcccta tcagtgatag
     2401 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     2461 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     2521 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     2581 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     2641 tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgagta ggcgtgtacg
     2701 gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     2761 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccccga
     2821 attcatggag ctgccaacaa agcctggcac cttcgacctg ggcctggcca catggagccc
     2881 ttccttccag ggggaaaccc accgggctca ggcacgccgc agggatgttg gcaggcagct
     2941 gcctgagtac aaggctgtgg tggtgggcgc cagtggcgtg ggcaagagtg cgctgaccat
     3001 ccagctgaac caccagtgct tcgtggagga ccacgacccc accatccagg attcctactg
     3061 gaaggagttg accctggaca gtggggactg cattctgaat gtgctggaca cagcagggca
     3121 ggccatccat agggccctgc gtgaccagtg cctggctgtc tgtgatggtg tgctgggcgt
     3181 cttcgctctc gatgacccct cgtctctgat ccagctgcag cagatatggg ccacctgggg
     3241 ccctcacccc gcccagcccc ttgtcctcgt gggcaacaag tgtgaccttg tgaccactgc
     3301 tggagatgct catgccgctg ctgcagccct cgcacacagc tggggggccc acttcgtgga
     3361 gacctcggcc aaaacacggc aaggcgtgga ggaggccttt tccctgctgg tccatgagat
     3421 ccagagggtc caggaggcca tggcgaagga gcccatggca aggtcctgta gggagaagac
     3481 ccggcaccag aaggccacct gccactgtgg ctgctctgtg gcctgagaat tcgatatcaa
     3541 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3601 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3661 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3721 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3781 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3841 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3901 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3961 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4021 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4081 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     4141 tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     4201 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     4261 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     4321 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     4381 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
     4441 agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     4501 gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag
     4561 cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct
     4621 agcatttcat cacatggccc gagagctgca tccggactcg agataacttc gtataatgta
     4681 tgctatacga agttattccg gactgtactg ggtctctctg gttagaccag atctgagcct
     4741 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4801 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4861 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
     4921 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4981 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5041 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5101 tgggaagaca atagcaggca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5161 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5221 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5281 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5341 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5401 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5461 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5521 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5581 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5641 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5701 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5761 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5821 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5881 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5941 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6001 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6061 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6121 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6181 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6241 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6301 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6361 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6421 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6481 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6541 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6601 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6661 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6721 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6781 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6841 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6901 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6961 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7021 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7081 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     7141 cgcgatgtac gggccagata ta
//
